Construction And Experimental Study On Micro - Environment Simulation Platform Of Bladder Cancer

Objective: To fabricate a microfluidic platform to reconstruct bladder cancer microenvironment with four types of cells co-culture(bladder cancer cell, fibroblast,macrophage and endotheliocyte). Based on this simulation platform, to explore the phenotype change of stromal cells and test drug sensitivity for bladder cancer cell treated with multiple types of chemotherapy drugs.Methods: Microfluidic technology was employed to fabricate this bladder cancer simulation platform. This platform consists of matrigel channel units, medium channel and four indirect contact culture chambers which enables to different types of cell interaction by soluble biological factors and metabolite. Immunofluorescence staining assay was employed to detect the differentially expressed α-SMA for fibroblast, CD105 for endotheliocyte, Arg-1 for macrophage between experimental group and control group.Bladder cancer cell was treated with multiple types of chemotherapy drugs by simulating clinical chemotherapeutical schemes to test the drug sensitivity.Results: Cell morphological observation showed that cells grew well in this device and T24 cells were observed having a tendency to form network structure at 3D culture condition by real-time observation. The macrophage migration tendency was observed by comparing the micrographs at different time points in this platform. Compared with control group, the effector factors α-SMA/CD105/Arg-1 were highly expressed in fibroblast/endotheliocyte/macrophage respectively. The death cell counts(/100) for bladder cancer T24 cells treated with several types of chemotherapy drugs were different based on this microfluidic platform.Conclusion: This study presented a platform to emulate bladder cancer microenvironment with multi-type cells co-culture in vitro. With this platform, stromal cells existed in bladder cancer microenvironment have the tendency to be activated.Chemosensivity test for bladder cell were carried out successfully on this microfluidic platform which also provided an excellent platform for further investigation of anti-drug mechanism for tumor cell.